目的 基于干扰素 γ（IFN-γ）/核因子 κB（NF-κB）/SNAIL 轴探讨复方蜥蜴散干预肿瘤细胞干性对胃癌顺 铂耐药的增敏提效作用机制。方法 （1）取对数生长期的 MKN45 细胞和 MKN45/DDP 耐药细胞，分别使用不同 浓度（0、0.2、0.4、0.8、1.6、3.2 μg·mL-1 ）的顺铂（DDP）处理细胞 48 h，采用 CCK-8 法测定细胞增殖能力及耐 药指数。采用无血清悬浮成球实验验证 MKN45 细胞及 MKN45/DDP 耐药细胞的肿瘤干细胞特性。（2）将对数生 长期 MKN45/DDP 细胞接种于 BALB/c 雄性裸鼠右前腋窝皮下，复制胃癌顺铂耐药裸鼠移植瘤模型。将模型复 制成功的裸鼠随机分为模型组、顺铂组、复方蜥蜴散组及联合组，每组 8 只。模型组灌胃等体积生理盐水；顺 铂组腹腔注射 0.002 g·kg-1顺铂，每周 2 次；复方蜥蜴散组灌胃 2.8 g·kg-1复方蜥蜴散水煎液，每天 2 次；联合 组灌胃复方蜥蜴散水煎液（2.8 g·kg-1 ，每天 2 次）的同时腹腔注射顺铂（0.002 g·kg-1 ，每周 2 次）；各组均连续干 预 4 周。称量肿瘤质量并计算肿瘤体积；采用 HE 染色法观察肿瘤组织病理变化；TUNEL 法测定肿瘤组织细 胞凋亡情况；免疫组化法检测肿瘤组织 Ki67、EpCAM、CD44 蛋白表达情况；Western Blot 法测定肿瘤组织多 药耐药相关蛋白 1（MRP1）、P-糖蛋白（P-gp）、EpCAM、CD44、IFN-γ、p65、p-p65、SNAIL 蛋白表达水 平。结果 （1）与 MKN45 细胞比较，0.8、1.6、3.2 μg·mL-1 顺铂组的 MKN45/DDP 耐药细胞存活率显著升高 （P＜0.05，P＜0.01）；MKN45 细胞的 IC50为 0.96 μg·mL-1 ，MKN45/DDP 细胞的 IC50为 2.26 μg·mL-1 ，MKN45/DDP 细胞的耐药指数为 2.36。与 MKN45 细胞比较，MKN45/DDP 耐药细胞的成球能力显著升高（P＜0.01）。（2）模型 组裸鼠的肿瘤细胞形态不规则，排列较紧密，细胞核大，可见病理性核分裂象。与模型组比较，各给药组胃癌 耐药 MKN45/DDP 细胞荷瘤裸鼠的肿瘤体积均显著缩小（P＜0.01），肿瘤质量均显著降低（P＜0.01）；肿瘤细胞 核分裂现象减少，细胞密度降低，排列较松散，可见细胞坏死象；肿瘤细胞凋亡率均显著升高（P＜0.01）；肿 瘤组织中 Ki67 阳性表达水平显著降低（P＜0.05，P＜0.01）；肿瘤组织中的干性标志物 EpCAM、CD44 及耐药 相关蛋白 MRP1、P-gp 的蛋白表达水平均显著降低（P＜0.05，P＜0.01）；肿瘤组织 p-p65/p65、IFN-γ、SNAIL 蛋白表达水平均显著降低（P＜0.01）。与顺铂组比较，复方蜥蜴散组及联合组胃癌耐药 MKN45/DDP 细胞荷瘤 裸鼠的肿瘤体积均显著缩小（P＜0.01），肿瘤质量均显著降低（P＜0.01）；肿瘤细胞凋亡率均显著升高（P＜ 0.01）；肿瘤组织中 Ki67 阳性表达水平显著降低（P＜0.05，P＜0.01），CD44、MRP1、P-gp、IFN-γ、SNAIL 蛋 白表达水平显著降低（P＜0.05，P＜0.01）；联合组裸鼠肿瘤组织中 EpCAM、p-p65/p65 蛋白表达水平显著降低 （P＜0.01）。结论 复方蜥蜴散能够有效抑制胃癌耐药 MKN45/DDP 细胞荷瘤裸鼠的肿瘤生长，抑制细肿瘤胞 增殖，促进其凋亡，可能与通过调控 IFN-γ/NF-κB/SNAIL 轴抑制肿瘤细胞干性，从而降低对顺铂的耐药性有关。

Objective To investigate the mechanism by which Compound Xiyi San（CXS） enhances cisplatin sensitivity in gastric cancer by interfering with tumor stemness via the interferon-γ（IFN-γ）/nuclear factor-κB （NF-κB）/SNAIL axis. Methods （1） MKN45 cells and MKN45/DDP cisplatin-resistant cells in the logarithmic growth phase were treated with different concentrations （0，0.2，0.4，0.8，1.6，3.2 μg·mL-1 ） of cisplatin （DDP） for 48 hours. Cell proliferation capacity and the resistance index were measured using the CCK-8 assay. The cancer stem cell characteristics of MKN45 and MKN45/DDP cells were validated using the serum-free suspension sphere formation assay.（2） MKN45/DDP cells in the logarithmic growth phase were subcutaneously inoculated into the right anterior axilla of male BALB/c nude mice to establish a cisplatin-resistant gastric cancer xenograft model. Successfully modeled mice were randomly divided into four groups （n=8 per group）：model group，cisplatin group，CXS group，and combination group. The model group received an equal volume of normal saline by gavage；the cisplatin group received intraperitoneal injections of 0.002 g·kg-1 cisplatin twice weekly；the CXS group received 2.8 g·kg-1 CXS aqueous extract by gavage twice daily；the combination group received both CXS aqueous extract （2.8 g·kg-1 ，twice daily） and cisplatin （0.002 g·kg-1 ，twice weekly）. All interventions continued for 4 weeks. Tumor mass was weighed，and tumor volume was calculated. Pathological changes in tumor tissue were observed by HE staining；apoptosis was detected by TUNEL assay；protein expression of Ki67， EpCAM， and CD44 in tumor tissue was determined by immunohistochemistry； and protein expression levels of multidrug resistance-associated protein 1 （MRP1）， P-glycoprotein （P-gp）， EpCAM， CD44， IFN- γ， p65， phosphorylated p65（p-p65），and SNAIL were measured by Western Blot. Results （1） Compared with MKN45 cells， the survival rates of MKN45/DDP cells in the 0.8，1.6，and 3.2 μg·mL-1 cisplatin groups were significantly increased （P＜0.05，P＜0.01）. The IC50 for MKN45 cells was 0.96 μg·mL-1 ，and for MKN45/DDP cells it was 2.26 μg·mL-1 ， resulting in a resistance index of 2.36 for MKN45/DDP cells. Compared with MKN45 cells，MKN45/DDP cells exhibited a significantly enhanced sphere-forming ability （P＜0.01）.（2） In the model group， tumor cells showed irregular morphology，tight arrangement，large nuclei，and pathological mitotic figures. Compared with the model group，all treatment groups showed significantly reduced tumor volume （P＜0.01） and tumor mass （P＜0.01） in MKN45/DDP xenograft nude mice；mitotic figures were reduced，cell density decreased，arrangement became looser，and necrotic cells were observed；tumor cell apoptosis rates were significantly increased （P＜0.01）；Ki67 positive expression levels in tumor tissue were significantly decreased （P＜0.05， P＜0.01）； protein expression levels of stemness markers EpCAM and CD44，and drug resistance-related proteins MRP1 and P-gp were significantly decreased （P＜0.05，P＜ 0.01）；and protein expression levels of p-p65/p65，IFN-γ，and SNAIL in tumor tissue were significantly decreased （P＜0.01）. Compared with the cisplatin group，the tumor volume and tumor mass of MKN45/DDP xenograft nude mice in the CXS group and the combination group were significantly reduced （P＜0.01）；significantly increased apoptosis rates （P＜0.01）； significantly decreased Ki67 positive expression （P＜0.05， P＜0.01）， and significantly lower protein expression levels of CD44，MRP1，P-gp，IFN-γ，and SNAIL （P＜0.05，P＜0.01）；the combination group also showed significantly decreased EpCAM and p-p65/p65 protein expression levels （P＜0.01）. Conclusion Compound Xiyi San effectively inhibits tumor growth，suppresses cell proliferation，and promotes apoptosis in nude mice bearing cisplatin-resistant MKN45/DDP gastric cancer xenografts. This effect may be related to the suppression of tumor stemness and subsequent reduction of cisplatin resistance through regulation of the IFN-γ/NF-κB/SNAIL axis.

R285.5

国家自然科学基金项目（82260916）；宁夏医科大学科学研究资助项目（XZ2023012）。